Western Blot Analysis of Apoptosis Regulators

Western blot analysis was performed as described previously (Wang et al., 2020 (link)). The following antibodies were used: Bcl-2 (#4223), Phospho-Bcl-2 (#2875), Bcl-xL (#2764), Bid (#2002), Bax (#5023), Bad (#9239), phospho-JNK (#4668), JNK (#9252), JNK1 (#3708), JNK2 (#9258), Cleaved Caspase-3 (#9664), PARP (#9532), phospho-c-Jun (Ser73) (#3270), phospho-c-Jun (Ser63) (#91952), c-Jun (#9165), PUMA (#4976), GAPDH (SC-47724). The antibodies were used at either 1:1,000 dilution or 1:2000 dilution. The GAPDH antibodies were purchased from Santa Cruz Biotechnology, and the rest of the antibodies were purchased from Cell Signaling Technology. The band intensity of the target protein was quantified using VisionWork LS image acquisition and analysis software (UVP).

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Chen S., Wang Q., Ming S., Zheng H., Hua B, & Yang H.S. (2022). Platycodin D induces apoptosis through JNK1/AP-1/PUMA pathway in non-small cell lung cancer cells: A new mechanism for an old compound. Frontiers in Pharmacology, 13, 1045375.

Publication 2022

GAPDH antibodies

Antibodies Bcl 2 C jun Caspase 3 Dilution Gapdh Protein target Puma Western blot analysis

Corresponding Organization :

Other organizations : Beijing University of Chinese Medicine, Markey Cancer Center

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression levels of the following proteins: Bcl-2, Phospho-Bcl-2, Bcl-xL, Bid, Bax, Bad, phospho-JNK, JNK, JNK1, JNK2, Cleaved Caspase-3, PARP, phospho-c-Jun (Ser73), phospho-c-Jun (Ser63), c-Jun, PUMA

control variables

GAPDH expression (used as a loading control)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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